Large ribosomal protein depletion impacts CB substructure and nuclear dimethylarginine profile. (A) Representative single CB immunofluorescent staining of coilin (magenta) and SMN (cyan) using STED microscopy in siNT, siRPL14, and siRPL24 cells. Two CBs per condition are shown. (B) Quantification of offset distance between CB and gems in HeLa cells after siRNA knockdown of control or ribosomal proteins for 72 h. The offset distance between CB and gems is calculated from coilin and SMN fluorescent intensity-weighted center of mass. The cyan line stands for the median offset distance in each sample. Number of CB-gem pairs measured: siNT = 356; siRPL14 = 460; siRPL24 = 229; siRPL6 = 152; siRPL17 = 178. P values are obtained by Kruskal–Wallis test followed by the Dunn test with Bonferroni correction. **, P < 0.01; ****, P < 0.0001. (C) Immunofluorescent images showing symmetric dimethylarginine (sDMA) signal in HeLa cell after respective siRNA knockdown. CB outline is shown as the magenta line. Scale bars = 5 μm. (D and E) Quantification of sDMA intensity in HeLa cell nucleus or CB after respective siRNA knockdown. The relative sDMA intensity is calculated by dividing mean DMA intensity in each nucleus (D) or CB (E) to the respective mean coilin intensity. Median of the relative DMA intensity is shown by the magenta (D) or cyan (E) line. Number of replicates = 4. P values are obtained in the same way as in B.